7^th International Conference and Exhibition on Analytical & Bioanalytical Techniques September 28-30, 2016 Orlando, Florida, USA

Dr. Steiner earned his Ph.D. in Analytical Chemistry with focus on the development and use of analytical instrumentation to explore a variety of topics involving health, environment, agriculture, and defense. Presently his research group is interested in applications that are focused on the qualitative discovery and quantitative directed analysis of bio-markers that can be correlated to a specific disease trait, state, and/or rate. Dr. Steiner has participated in obtaining more than 20 grants, the publication of more than 30 publications, and the presentation of more than 75 presentations

The advancement of hyphenated instrumentation from a variety of scientists working in the public and private sectors of research and development in the form of gas chromatography (GC) being coupled to mass spectrometry (MS) has continued to develop into a powerful multi-dimensional hyphenated instrumental technology for the use in a wide assortment of analytical and bioanalytical techniques. Examples of this development can be seen in the analysis of drugs, metabolites, pesticides, chemical warfare agents, food ingredients, medications, fuels, and etcetera and/or in the main category of volatile and semi-volatile organic compound analysis in fields such as forensic, toxicology, environment, defense, food and beverage, pharmaceutical, petrochemical, and etcetera. This advancement in hyphenated GC-MS instrumentation was initially and still is driven by the need for a more comprehensive analytical and bioanalytical technique that can accurately and precisely discriminate targeted and untargeted analytes from higher complexity sample mixtures in a sensitive and selective way from within a concise window of time. With this in mind, this presentation which is based upon a recent editorial, briefly attempts to highlight some of the current trends in hyphenated GC-MS instrumentation available today and their respective contributions to the field of analytical and bioanalytical techniques.

Lihua GU is a PhD student in Shanghai University of Traditional Chinese Medicine. She is engaging in the research and application of HPTLC, TLC-bioautographic methods and quality control standard in TCM. She has published more than 10 academic papers.

TLC bioautography is an assay that combines chromatographic separation and in situ determination of biological activity. It has advantages over other screening methods for its ability to screen mixtures (a wide solubility range of compounds) simultaneously. In addition, it can reduce the cost and time of screening. TLC autobiography assays have been established for screening compounds for antimicrobials, antioxidants (free-radical scavengers) and enzyme inhibitors. In this presentation the application of TLC bioautographic techniques on enzymatic reaction will be reviewed. An example of the development of a novel TLC-bioautographic method on detection of dipeptidyl peptidase IV (DPP IV) inhibitors from plant extracts will be described. The focus of the example will be on how reaction products via derivatisation can facilitate such screening technology. In this case the enzyme (DPP IV) hydrolyzes substrate (Gly-Pro-p-nitroaniline) into p-nitroaniline (pNA), which diazotizes with sodium nitrite, and then reacts with N-(1-naphthyl) ethylenediamine dihydrochloride in turn to form a rose-red azo dye which provides a rose-red background on the TLC plates. The DPP IV inhibitors showed white spots on the background as they blocked enzymolysis of the substrate to produce pNA. The method was validated with respect to selectivity, sensitivity, linearity, precision, recovery, and stability after optimizing key parameters including plate type, time and temperature of incubation, concentration of substrate, enzyme and derivatization reagents, and absorption wavelength. One natural compound harmine was isolated and identified to be a potential DPP IV inhibitor in nine medicinal herbs by this method.

Young jin Koh graduated from Department of Chemistry, Duksung women’s University and obtained her master degree from Department of Analytical Chemistry, Seoul national University in South Korea. At present, she is a researcher at Agency for Defense Development in South Korea

A Raman spectroscopy is a spectroscopic technique based on inelastically scattered light and is widely used for the analysis of materials. The Raman spectrometer is very effective in detecting liquid chemicals on surfaces on the move. Due to advanced ultraviolet light source technology, the Raman spectrometer based on ultraviolet laser has been actively developed, recently. In case of using below 250 nm wavelength as a deep UV light source, it is easy to measure the Raman spectra by spectrally separating the Raman and the fluorescence area. In this research, we describes about analysis of Raman spectra for chemicals on various deep UV wavelength and compared with Raman spectra according to laser wavelengths.

Dr. Young-Su Jeong completed his PhD from Korea Advanced Institute for Science and Technology (KAIST) in 2012. He is a senior researcher of Agency for Defense Development (ADD), ROK. He has published more than 18 papers in reputed journals

A Raman Agent Monitoring System (RAMS) is a technique which exploits Raman scattering to provide stand-off detection and identification of toxic chemicals such as chemical warfare agents and toxic industrial chemicals deposited on the ground surfaces. The RAMS collects counts on photons emitted by an agent on a surface through a telescope and displays the result on a spectrograph. Raman scattering occurs when light interacts with a molecule. The molecule vibrates and scatters wavelength shifted light. The wavelength shifts and their associated intensities are dependent on the size, shape, and bond strength of the molecule creating, thus, a distinct spectral signature for the associated chemical. In this research, we measured toxic chemicals with a RAMS, a 248 nm mobile Raman spectrometer, and analyzed Raman spectral signatures of Raman signals of that chemicals. These results can assist in selecting unique molecular markers for each chemicals to allow real-time detection and identification of them in ground environments in moving.

JQ Zhang has completed his MD at 2013 from Nanjing University of Chinese Medicine. She is dean of the College of pharmacy, Hainan Medical University. She has published more than 25 papers in reputed journals and has been serving as an editorial board member of repute

Galangin (3,5,7-trihydroxyflavone) is present in high concentrations in Alpinia officinarum Hance. It shows multifaceted in vitro and in vivo biological activities. The number and position of hydroxyl groups in this molecule play an important role in these biological activities. In order to systemic clarify the exposure of galangin after oral and intravenous administration to rats, two LC-MS/MS methods were developed and validated and successfully applied to analyze the parent drug molecules and aglycones liberated from plasma samples via β-glucuronidase hydrolysis. Our major findings were as follows: (1) The routes of administration showed significant influences on the systemic exposure of galangin and its metabolites. (2) Galangin was preferentially glucuronidated after p.o. dosing but sulfated after i.v. medication. (3) Kaempferol conjugates were detected demonstrating that oxidation reaction occurred; however, both glucuronidation and sulfation were more efficient. (4) Oral bioavailability of free parent galangin was very low. The results show that systemic exposure to galangin and its metabolites was different in rat plasma between oral and intravenous administration. Further research is needed to characterize the structures of galangin conjugates and to evaluate the biological activities of these metabolites.

Ling Huang has completed her PhD at the age of 34 years from School of Pharmacy of SUN YAT SEN University. She has published more than 10 papers in reputed journals and has been serving as an editorial board member of repute. (Up to 100 words)

Feng-Liao-Chang-Wei-Kang (FLCWK) is a traditional Chinese patent medicine that mainly consists of Daphniphyllum calycinum roots and Polygonum hydropiper. As a complex containing several types of flavonoids, FLCWK has potential effects on the drug metabolism enzyme P450 3A4(CYP3A4) and nuclear receptors. This study aims to explore the effects of FLCWK on CYP3A1(CYP3A4′s homolog in rats) in rats and to determine whether FLCWK could participate in the processes of hPXR- and hCAR-mediated transactivation of CYP3A4. The effects of FLCWK on CYP3A1 mRNA, protein expression and catalytic activity levels in Sprague-Dawley(SD) rat liver tissues were detected using real-time PCR, western blotting and high- performance liquid chromatography (HPLC) assays. The effects of hPXR and hCAR on CYP3A4 transcriptional activity were examined using luciferase reporter gene assays; the futher study of FLCWK on the CYP3A4 gene expression mediated by PXR pathway was investigated by transient transfection of PXR siRNA . This study found that FLCWK could significantly increase the CYP3A1 mRNA gene and protein expression levels and CYP3A1 activity in SD rats. In PXR-CYP3A4 co-transfected cells, FLCWK could significantly induce CYP3A4 luciferase activity mediated by PXR. PXR-knockdown (transfected with siPXR construct) decreased the CYP3A4 mRNA level than in the control cells transfected with corresponding vector. Taken together, these findings suggest that FLCWK could significantly upregulate CYP3A4 levels via the PXR-mediated pathway. This effect should be taken into consideration to predict any potential drug-drug interactions between FLCWK and other co-administered drugs. (Up to 250 words)

Ghazale Daneshvar Tarigh received her PhD degree in analytical chemistry from University of Tehran, Iran in 2015. She received her bachelor's degree (B.Sc.) in pure chemistry at the University of Zanjan, in 2003. She got her master's degree (M.Sc.) under the direction of Prof. Yadollah Yamini at TMU and Prof. Ali Jabbari at KNTU in 2009. Her field of interest is the development of new extraction technologies, with an emphasis on miniaturized sample preparation methods and separation techniques.

L-threonine (L-thr) as a general chiral selector anchored on the surface of magnetic multi wall carbon nanotube (MMWCNT) was prepared using an in situ electrostatic adsorption and studied as a new magnetically chiral selector for the separation of chiral DL-mandelic acid (DL-MA) as a model sample. By varying the pH, DL-MA was adsorbed on the surface of magnetic chiral selector through hydrogen bonds. It is recognized that MMWCNT with chiral ligands on its surface simultaneously possess both magnetic property and direct chiral recognition ability. The successful immobilization of L-thr onto the surface of MMWCNT was confirmed by infrared spectra (FT-IR), X-ray diffraction patterns (XRD) and transmission electon microscopy (TEM). The FT-IR and mass spectra of supernatant and elution solutions also confirmed the immobilization of L-thr onto the surface of MMWCNT. The analysis results of specific rotation, HPLC and ultraviolet–visible spectroscopy reveal that the L-thr-MMWCNT show stronger complexation of (+)-enantiomer than (-)-enantiomer. The functional magnetic nanotubes were easily separated from the racemic solution using an external magnetic field which demonstrated its feasibility of recycling the adsorbent. All processes including in situ immobilization, enantioseparation (enantioenrichment) and magnetic separation were done by single process in a short time (only 10 min).

Wang Wei, associate professor, Master of Electronic Engineering, graduated from Jilin University. The research is mainly engaged in analytical instrumentation design. Till now he has designed and developed a variety of analytical detection equipments. He obtained progress Award of Jilin Province Science and Technology in nano-structured composites and analysis instrumentation aspects in 2008.

The piezoelectric ceramic positioning system is considered to be the high-precision positioning device in the electrochemical imaging detector, which can achieve the nanometer scale displacement resolution. This paper describes in detail the principle of output displacement measuring for piezoelectric ceramic by the resistance strain gauge. In addition, by employing the self-built drive circuit and the displacement detecting circuit, integrated piezoelectric ceramic resistance strain gauge sensor has been investigated under both open-loop and closed-loop control, respectively. Under the closed-loop control testing, with a classic PID algorithm was applied. Test results show that, the piezoelectric ceramics in open-loop control exhibits disadvantages of serious non-linearity, hysteresis and creep resistance. However, these issues have been resolved under closed-loop control.

Dr. Anna Gajda has completed her PhD in 2014 at Department of Pharmacology and Toxicology, NVRI, Pulawy, Poland. She is responsible for antimicrobial residues determination in food of animal origin by chromatographic techniques (LC-MS/MS, HPLC-FLD/UV). She participates in realization of National Residues Control Plan in Poland. She is engaged in development a new analytical methods for the detection of veterinary drugs in food products. She has published more than 23 papers in reputed, international journals.

The misuse and unresonable administration of antibiotics in swine may lead to residues occurrence in products of animal origin. The main material for antibacterials determination is tissue of animals. Alternative for the post-mortem residues control is oral fluid analysis, as a non-invasive, ante-mortem method for the antibiotics detection. Oxytetracycline is one of the most widely used tetracycline in swine treatment. To proved and demonstrated the utility of oral fluid for the detection of this compound, oxytetracycline was administered by intramuscular injection to swine and oral fluid samples were analysed. For the measurement of oxytetracycline in oral fluid, a liquid chromatography – tandem mass spectrometry (LC-MS/MS) method was developed. The extraction was carried out with 10% trichloracetic acid. Samples were cleaned up by filtration using PVDF filters. Chromathografic separation was achieved on a Luna C18 analytical column using mobile phase consisting of acetonitrile and 0.1 % formic acid in gradient mode. During validation a good linearity was observed (r > 0.99). The recoveries were in rhe range of 90 - 105%. The validation results showed good accuracy with a good RSD, less than 10.0% for repeatability and less than 15% under within-laboratory reproducibility. The procedure was satisfactory sensitive with detection limit LOD = 2 µg/kg and limit of quantification LOQ = 5 µg/kg. The presence of oxytetracycline in oral fluid up to 21 days after IM injection indicate, that this medium has the potential to be an effective way to test the antibiotic residues in live animals.

Tomasz Bladek is a research assistant at the Department of Pharmacology and Toxicology, National Veterinary Research Institute in Pulawy, Poland. His major research interests lie in the field on the development of new analytical methods for determination of antibiotic residues in food matrices such as muscle, milk, kidney, liver, eggs and honey. He has published several IF papers on antibiotic residues and has been serving as a reviewer for analytical journals

A liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method for determination and quantification of sulfonamides, tetracyclines, macrolides, fluoroquinolones, streptomycin, lincomycin and trimethoprim resideues in honey has been developed. Prior to instrumental analysis, sample preparation technique involved sample preparation technique involved extraction with ammonium acetate buffer followed by solid-phase extraction cleanup using polymeric reversed phase sorbent. The chromatographic separation was performed on C18 column with gradient elution mode. Mass spectral acquisitions were performed under selective multiple reaction monitoring mode by triple quadrupole mass spectrometry detector. The method has been characterized by providing the parameters of precision, accuracy, specificity, decision limit (CCα) and detection capability (CCβ) according to Commission Decision 2002/657/EC. The calibration curves were linear in the range of 5-100 μg/kg. The recoveries ranged from 86 to 104%. The repeatability was below 11.3% and within-laboratory reproducibility was lower than 16.1%. Calculated CCα and CCβ values ranged from 5.8 to 7.6 μg/kg and from 6.9 to 9.9 μg/kg, respectively. The developed method fulfils the criteria for confirmatory methods and thanks to its labour efficiency, it may be used also for screening purposes.

Pharmaceutical honours degree with merit from the Programa Universidade Para Todos (PROUNI) rated best pharmacy student at the Centro Universitário Central Paulista, São Carlos, Brazil (2009). Master in Pharmaceutical Sciences from the Universidade Estadual Paulista, Brazil (UNESP) Araraquara, conducted research on the quality control of drugs and medicines, with emphasis on development and validation of analytical methods. Currently conducting research with the antimicrobial ertapenem sodium for the development of PhD research at the Faculdade de Ciências Farmacêuticas, Brazil and PhD exchange student at the University of Ghent – Ugent, Belgium and is funded by FAPESP financing agency.

Ertapenem sodium is the polar and ionizable compound. Thus, the most suitable for such drug is to develop a method for Hydrophilic Interaction Chromatography (HILIC). On the other hand, the RP-HPLC is considered the analysis techniqueis more used for quantification of substances and new column are often introduced for compounds separation. Two new analytical methods have been developed. The RP-HPLC method was performed on a Waters LC system, with an Agilent™ Zorbax Bonus-RP (4.6 x 150 mm, 5 μm) column and the mobile phase was water: ethanol 80:20 v/v. While the HILIC method was performed on an ultra-high performance liquid chromatography Shimadzu with stationary phase PhenomenexTM HILIC Kinetex (4.6 x 100 mm, 2.6 µm) column and the mobile phase was acetonitrile:water 88:12 v/v. In both methods was added 0.1 % formic acid in mobile phase and was performed isocratically. The validation parameters were in accordance with ICH specifications and demonstrated accuracy, precision, selectivity, robustness and linearity. Analytical curves was constructed and was evaluated by ANOVA (y = 23043x - 38525, R2= 0.9999 RP-HPLC and y = 29928x - 547879, R2= 0.9994 HILIC). The robustness was evaluated by making parameters small alterations simultaneous and was evaluated by the Plackett- Burman. The results show that the method validity is maintained. The methods here described have economic advantage as well as features an eco-friendly focus. This work was developed with environmental conscience, always looking to minimize the possible generated organic waste, which is of utmost importance to the environment.

Graduated in Pharmacy from the Centro Universitário de Votuporanga (2010). Master in Biotechnology from the Universidade Estadual Paulista - UNESP (2013). He is currently a doctoral student (PhD) in Pharmaceutical Sciences program of the School of Pharmaceutical Sciences of Araraquara - FCFAr (UNESP) and is funded by FAPESP.

The prospect of new analytical methods, the improvement and validation of existing methods, bring numerous benefits to the pharmaceutical industry, for the purpose to save as much in costs as the analysis time. Cefepime hydrochloride (CEF), an antimicrobial agent β-lactam belonging to the group of fourth generation cephalosporins, is a semi-synthetic product which has activity against several Grampositive and Gram-negative aerobic bacteria. The objective of this study was performed a statistical analysis of average contents of CEF obtained by reversed-phase high-performance liquid chromatography (RP-HPLC), microbiological assay (turbidimetric test) and UV method, using analysis of variance (ANOVA). The RP-HPLC method was performed on a C18 column (250 mm x 4.6 mm) maintained at room temperature. The mobile phase consisted of water : absolute ethanol (45:55, v/v) at a flow rate of 0.5 mL min-1, using UV detection at 258 nm. For performing the turbidimetric assay, Staphylococcus aureus ATCC 6538 IAL 2082 was used as the test microorganism and the culture medium chosen was the Casoy broth. Was used temperature control (35.0 °C ± 2.0 °C) and incubated for4 hours in shaker. The readings of the results were made in spectrophotometer at 530 nm. The UV method was realized using the equipment Spectrophotometer UV Shimadzu. The absorption was obtained at a wavelength of 258 nm. The results through ANOVA showed a significant difference between the methods proposed for the 5% significance level. Thus, the methods are not equivalent and should be used in conjunction in the quality control analysis of CEF.